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Objective:To investigate the application effect of case-based learning (CBL) combined with flipped classroom in the teaching of experimental diagnostics in the integrated course of Diagnosis and Treatment Fundamentals. Methods:The cluster random sampling method was used to select the class of 2019 in the eight-year program and the class of 2020 in the five-year program, with the major of stomatology in Air Force Medical University. The 24 students in the observation group received CBL combined flipped classroom, and the 37 students in the control group received traditional teaching. The two groups were compared in terms of theoretical assessment score, classroom assessment score, comprehensive ability, self-learning ability, and degree of satisfaction with teaching. SPSS 22.0 was used for the t-test. Results:The observation group had a significantly higher theoretical assessment score than the control group [(74.88±3.46) vs. (71.89±4.45), P<0.05]. The observation group had significantly better scores of practical skill assessment than the control group ( P<0.05). Compared with the control group, the observation group had significantly better scores of comprehensive ability and self-learning ability ( P<0.05). The observation group had significantly better scores of satisfaction with teaching than the control group ( P<0.05). Conclusion:The application of CBL combined with flipped classroom in the teaching of experimental diagnostics in the integrated course of Diagnosis and Treatment Fundamentals can improve theoretical knowledge, practical skills, comprehensive quality, and satisfaction with teaching among students, and therefore, it holds promise for application in teaching.
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【Objective】 To investigate treatments of discrepancy presented in blood typing and cross-matching test caused by anti-I antibody and the difference between autoanti-I antibody and alloanti-I antibody. 【Methods】 38 cases of I-positive antibody in our hospital from January 2016 to July 2019 were selected as the research subjects. The irregular antibodies screening and identification were performed by adopting the anti-human globulin and saline test tube method, and the blood transfusion effect was evaluated. 【Results】 37 cases of autoanti-I antibody and 1 case of alloanti-I antibody, with specificity produced by an individual with a rare i blood group, were identified. 34 cases contained I-positive antibody and 4 contained I-positive antibody combined with alloantibodies. At 4 ℃, most of the anti-I titers were between 32 and 512, 2 cases were more than 1 024. After the RBCs were washed with 37℃ normal saline and cold absorbed at 4℃, the cross-matching tests were matched and 37 cases of blood transfusion were all effective except for one case. After performing the same treatment on i adult red blood cells and adding I antigen-negative cord blood cells, the result was correct to be B type. The titer of IgM alloanti-I antibody in this adult was 256, and autotransfusion was preferred. 【Conclusion】 Patients with anti-I antibody, reactive at low temperature, should be treated with warm and slow transfusion under close monitoring. Autotransfusion is, in principle, beneficial to adult i blood group patients producing alloanti-I antibody. If i blood patients suffered from massive blood loss without suitable blood resource available, the elderly i blood donors were preferred, and plasmapheresis may also be an alternative to remove anti-I temporarily.
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Objective To investigate the relationship between tenascin-C (TNC) and acute rejection (AR) early after liver transplantation. Methods Six Brown Norway (BN) rats and 16 Lewis rats were divided into the AR group (Lewis→BN, 6 donors and 6 recipients) and control group (Lewis→Lewis, 5 donors and 5 recipients). The transplant liver tissues from rats in two groups were subjected to pathological examination. The rejection activity index (RAI) was evaluated by Banff schema. The expression of TNC proteins in the transplant liver tissues were determined by immunohistochemical staining and Western blot. The expression level of serum TNC was detected by enzyme-linked immune absorbent assay (ELISA), and the correlation between TNC and RAI score was analyzed. Results Pathological examination of the transplant liver at 7 d after liver transplantation showed that the RAI score in the AR group was higher than that in the control group. Immunohistochemical staining results found that the distribution of TNC positive cells of the transplant liver in the AR group was more than that in control group at postoperative 7 d. Western blot showed that the relative expression level of TNC protein in the AR group was significantly higher than that in the control group at 7 d after liver transplantation (t=5.112, P=0.007). ELISA results revealed that the serum TNC expression level in the AR group was significantly higher than that in the control group at 7 d after liver transplantation (t=3.152, P=0.012). The serum TNC expression level was positively correlated with the RAI score (r=0.790 9, P=0.004). Conclusions The expression level of TNC is associated with AR after liver transplantation. TNC may become a novel target for diagnosis and treatment of AR early after liver transplantation.
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Objective To study the changes of intracellular calcium ion concentration in pulmonary artery smooth muscle cells (PASMCs) of hypoxic-induced persistent pulmonary hypertension (PPH) induced by calcium-sensitive receptor (CaSR) in a newborn mouse model.Method Ninety-six newbom C57BL/6 mice were randomly divided into control group,PPH group,PPH + agonist group and PPH + inhibitor group,with 24 mice in each group.The PPH model was induced by 12％ oxygen for 14 days.In the beginning,intraperitoneal injection of CaSR agonist (GdCl3) and CaSR inhibitor (NPS2390) were performed to mice in PPH + agonist group and PPH + inhibitor group respectively daily.After 14 days of modeling,pulmonary artery smooth muscle cells (PASMCs) of all four groups were cultured in vitro.Changes of Ca2+ fluorescence intensity in PASMCs of the four groups were detected by laser confocal microscope continuously.Result The ratio of pulmonary small vascular wall thickness to the vascular diameter and right ventricle/left ventricular thickness in PPH group were greater than those in the control group [(21.1％ ±1.8％) vs.(27.0％ ±0.9％),(0.62 ±0.22) vs.(0.83±0.45)],the differences were statistically significant (P ＜ 0.05),which imply that PPH mouse model was constructed successfully.The average Ca2+ fluorescence intensity in PASMCs of control group,PPH group,PPH + agonist group and PPH+ antagonist group were 122.5 ± 3.0,2 058.8 ±46.3,2 286.6 ±51.4 and 1 134.8 ± 8.5,respectively.The average Ca2+ fluorescence intensity in PASMCs of the PPH group,PPH + agonist group and PPH + antagonist group was higher than that of the control group respectively,the average Ca2+ fluorescence intensity in PASMCs of PPH group was higher than that of PPH + antagonist group,the differences were statistically significant (P ＜ 0.05).Whereas the difference of average Ca2 + fluorescence intensity in PASMCs of PPH group and PPH + agonist group was of no statistical significance (P ＞ 0.05).Conclusion CaSR may be involved in the occurrence and development of hypoxic-induced PPH in neonatal mice by affecting the intracellular Ca2+ concentration in pulmonary artery smooth muscle cells.
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Objective To evaluate the antibacterial effect of mononuclear cells(MCs)in the liver lavage solution. Methods For in vitro experiment, MCs were col ected from the liver lavage solution of SD rats and divided into the supplement of interleukin(IL)-15 and non-supplement groups. The MCs were co-cultured with Pseudomonas aeruginosa (P. aeruginosa)for 4 h and then the supernatant was collected and MCs were lysed. The bacterial load in the lysate was detected after LB plate culture. The levels of interferon(IFN)-γand tumor necrosis factor(TNF)-αin the supernatant were measured by enzyme-linked immune absorbent assay(ELISA). For in vivo experiment, 40 SD rats were administered via tracheal injection of P. aeruginosa solution at a dose of 1×109 CFU/mL and randomly divided into four groups(n=10).In the control group, physiological saline was given via gavage. In the immunosuppression group, tacrolimus(FK506)was delivered via gavage. In the MC group, MCs at a dose of 1.0×108 was given via intravenous injection after use of FK506. In the IL-15 pretreated-MC group, IL-15 pretreated-MCs at a dose of 1.0×108 were administered via intravenous injection after application of FK506. The lavage solution of pulmonary alveolus and the rat lung tissue were col ected. The bacterial load was detected after LB plate culture. The expression of IFN-γand TNF-αin the pulmonary alveolus and lung tissue were measured by ELISA and Western blot. Results Compared with MCs alone, IL-15 pretreated-MCs exhibited significantly higher antibacterial capability in vitro. The CFU was 35%of untreated MCs. The synthesis and release capabilities of IFN-γandTNF-αweresignificantlyenhanced.Comparedwiththecontrolgroup,thequantityofimmunecelsinthelungtissuewas decreased and the bacterial load in the lung tissue and the lavage solution of pulmonary alveolus was significantly increased, whereas the expression levels of IFN-γand TNF-αtended to decline in the immunosuppression group. Administration of IL-15 pretreated-MCs significantly enhanced the quantity of immune cel s in the lung tissue, decreased the bacterial load and increased the secretion of IFN-γand TNF-α. Conclusions MCs in the liver lavage solution exhibit favorable antibacterial activity.Underimmunosuppressioncondition,thedefensecapabilityofthehostagainsttheopportunisticpathogenicbacteria is significantly enhanced.
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Objective:To study the construction of matrix metalloproteinase 3 (MMP3)biosensor vector,and to illuminate the activated process of MMP3 in the living cells.Methods:The ECFP-MMP3-YPet biosensor vector anchored on cellular surface was constructed and identified.The MMP3 biosensor was transfected into the 293T cells.The transfection efficiency was observed 24 h after transfection.The flurorescence resonance energy transfer (FRET )-based MMP3 biosensor was observed by inversion fluorescence microscope. Results:The MMP3 biosensor vector was successfully constructed.The length of MMP3-YPet identified by double enzyme digestion and PCR was about 780 bp.The transfection efficiency of MMP3 biosensor was about 40%,and which was evenly presented in cytoplasm of 293T cells.And the FRET ratio of MMP3 biosensor was decreased after stimulation with uPA on the 293T cells. The FRET ratio reached its minimum about 30 min later. Conclusion:The MMP3 biosensor can sensitively and reliably monitor the MMP3 activation in living cells.
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Objective To construct the eukaryotic expression vector urokinase-type plasminogen activator (uPA) biosensor which was the composition of the fusion protein enhanced cyan fluorescent protein-uPA (substrate)-yellow fluorescent protein variant (ECFP-uPA substrate-linker-YPet).Methods By the template Src-biosensor, the YPet primers were designed by Primer Premier 5.0 software,and the restriction enzyme sites,uPA substrate gene sequence and linker were added in its 5′ end. With the intermediate vector pDMTM-18T, an eukaryotic expression vector which contained a fusion protein of ECFP-uPA substrate-linker-YPet was constructed by genetic engineering.Then the uPA biosensor was transfected into 293T cells.The transfection efficiency and expression of fusion proteins were observed after 24 h.Fluorescence resonance energy transfer (FRET)was observed by the inversion fluorescence microscope and measured by the MetaFlour FRET 4.6 software. Results The uPA biosensor vector was confirmed by the fragment of PCR and double restriction enzyme digestion.The transfection efficiency was nearly 40%.The immunofluorescence detection results displayed that uPA biosensor fusion protein expressed in the 293T cells membrane and the FRET of uPA biosensor in the living 293T cells was observed after incubation with the recombinant human uPA (rhuPA).Conclusion uPA biosensor is successfully constructed and it could be used as a molecular probe to study the temporal and spatial variation of uPA in living cells.
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The transforming growth factor-β1 (TGF-β1)/Smad3 signal pathway is related to mutiple physiological and pathological generation mechanism of human being. Up to date, however, the spacial and time information on the phosphorylated Smad3 is still unclear. In this study, the process of Smad3 phosphorylation was observed under the physiological state in the living cells. Firstly, the ECFP-Smad3-Citrine (Smad3 biosensor) fusion protein expression vector was constructed and identified. Then the Smad3 biosensor was transfected into 293T cells. The transfection efficiency and the expressions of fusion proteins were observed in 24 hours. Thirdly, Smad3 biosensor flurorescence resonance energy transfer (FRET) was observed with the inversion fluorescence microscope and measured by the MetaFlour FRET 4. 6 software. Smad3 biosensor transfection efficiency was nearly 40% and the fusion protein was seen under the fluorescence microscope. The FRET ratio of Smad3 biosensor in living 293T cells was decreased after 10 minutes incubation with the ligand of TGF-β1. The period of decreasing CFP and enhancing Citrine signals was about 300 seconds. With the technology of FRET, the TGF-β1/Smad3 signal pathway could be real time monitored dynamically under the physiological condition in living cells.
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Humanos , Transferencia Resonante de Energía de Fluorescencia , Vectores Genéticos , Células HEK293 , Microscopía Fluorescente , Fosforilación , Transducción de Señal , Proteína smad3 , Metabolismo , Programas Informáticos , Transfección , Factor de Crecimiento Transformador beta1 , MetabolismoRESUMEN
Objective To investigate the clinical significance of direct antiglobulin test(DAT) in patients with chronic hepatitis B.Methods Column agglutination technique(CAT) and conventional tube technique(CTT) were used to detect red blood cell(RBC) antibodies in a total of 162 samples,including 50 cases of asymptomatic carriers,42 cases of active phases of chronic hepatitis B(CHB) patients,52 cases of severe hepatitis and 18 healthy individuals.Results The RBC count and hemoglobin(Hb) in patients with severe hepatitis were significantly lower than those in asymptomatic carriers,CHB patients and healthy individuals(P
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Objective:To compare the pharmacokinetics and relative bioavailability between matrine sustained release tablet,matrine conventional capsule and matrine injection. Methods: Dogs were given single oral dose and multiple doses of matrine sustained release tablet, capsule or injection in a randomized crossover way. Matrine concentrations in dog plasma were determined by RP-HPLC method. Results: The results showed that the t max and c max were 300 min and (5.088?0.490) ?g/ml for matrine sustained release tablet, (85?12) min and (6.360?0.215) ?g/ml for the conventional capsule, and 10 min and (6.500?0.404) ?g/ml for the injection. The relative bioavailability of the sustained release tablet was (153.7?9.4)% compared with that of the conventional capsule; the absolute bioavailability of the sustained release tablet was (73.5?14.2)% compared with that of the injecetion. The in vivo absorption rate of the sustained release tablet was significantly correlated with the in vitro release rate(r=0.981 2,P
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Objective:To investigate the effect of Siwutang on promoting hematopoietic function of mouse splenocytes. Methods:Effect of Siwutang on concanavalin A (Con A)-primed mice splenocytes production of IL-3 and IL-2 was studied by 3H-TdR incorporation and dot blot hybridization. Results: Siwutang significantly enhanced Con A-primed mice splenocytes production of IL-3 and IL-2(P